Phospho-ATF2 (T71) Recombinant Rabbit Monoclonal Antibody [SC05-90]
Catalog# ET1610-30
Phospho-ATF2 (T71) Recombinant Rabbit Monoclonal Antibody [SC05-90]
-
WB
-
IF-Cell
-
IF-Tissue
-
IP
-
IHC-P
-
Human
-
Mouse
-
Rat
-
unconjugated
Overview
Product Name
Phospho-ATF2 (T71) Recombinant Rabbit Monoclonal Antibody [SC05-90]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Thr71 of Human ATF2 aa 58-94 / 505.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IP, IHC-P
Molecular Weight
Predicted band size: 55 kDa
Positive Control
Hela, NIH/3T3, SH-SY5Y, rat brain tissue, human prostate tissue, human uterus tissue, mouse colon tissue, human cervix tissue, human skin tissue, human breast tissue, human esophagus tissue, human placenta tissue, mouse large intestine tissue, Hela cells treated with 250ng/mL anisomycin for 30 minutes.
Conjugation
unconjugated
Clone Number
SC05-90
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:1,000
-
IF-Cell
-
1:50-1:200
-
IF-Tissue
-
1:50-1:200
-
IHC-P
-
1:100-1:800
Target
Function
Eukaryotic gene transcription is regulated by sequence-specific transcription factors which bind modular cis-acting promotor and enhancer elements. The ATF/CREB transcription factor family binds the palindromic cAMP response element (CRE) octanucleotide TGACGTCA. The ATF/CREB family includes CREB-1, CREB-2 (also designated ATF-4), ATF-1, ATF-2 and ATF-3. This family of proteins contain highly divergent N-terminal domains, but share a C-terminal leucine zipper for dimerization and DNA binding. ATF-2 forms homodimers and heterodimers with c-Jun to initiate CRE-dependent transcription. Phosphorylation of ATF-2 at Thr 69 and Thr 71 by stress-activated kinases is necessary for transcriptional activation. Myc also induces phosphorylation of ATF-2 at Thr 69 and Thr 71 to prolong the half-life of ATF-2. ATF-2 also functions as a histone acetyltransferase (HAT) by specifically acetylating histones H2B and H4 in vitro. The gene encoding human ATF-2 maps to chromosome 2q31.1.
Background References
1. Namachivayam, K. et al. 2015. All-Trans Retinoic Acid Induces TGF-β2 in Intestinal Epithelial Cells via RhoA- and p38α MAPK-Mediated Activation of the Transcription Factor ATF2. PloS one. 10: e0134003.
2. Lindaman, LL. et al. 2013. Phosphorylation of ATF2 and interaction with NFY induces c-Jun in the gonadotrope. Molecular and cellular endocrinology. 365: 316-26.
Sequence Similarity
Belongs to the bZIP family. ATF subfamily.
Tissue Specificity
Ubiquitously expressed, with more abundant expression in the brain.
Post-translational Modification
Phosphorylation of Thr-69 by MAPK14 and MAPK11, and at Thr-71 by MAPK1/ERK2, MAPK3/ERK1, MAPK11, MAPK12 and MAPK14 in response to external stimulus like insulin causes increased transcriptional activity. Phosphorylated by PLK3 following hyperosmotic stress. Also phosphorylated and activated by JNK and CaMK4. ATM-mediated phosphorylation at Ser-490 and Ser-498 stimulates its function in DNA damage response. Phosphorylation at Ser-62, Thr-73 and Ser-121 activates its transcriptional activity. Phosphorylation at Thr-69 or Thr-71 enhances its histone acetyltransferase (HAT) activity.
Subcellular Location
Mitochondrion outer membrane, Nucleus, Cytoplasm.
Synonyms
Activating transcription factor 2 antibody
Activating transcription factor 2 splice variant ATF2 var2 antibody
ATF 2 antibody
Atf-2 antibody
Atf2 antibody
ATF2 protein antibody
ATF2_HUMAN antibody
cAMP Response Element Binding Protein 2 antibody
cAMP response element binding protein CRE BP1 antibody
cAMP response element-binding protein CRE-BP1 antibody
ExpandImages
-
☑ Cell treatment (CT)
Western blot analysis of Phospho-ATF2 (T71) on HeLa cell lysates treated with or without untreated anisomycin(250ng/ml 30min) with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/1,000 dilution.
Lysates/proteins at 20 µg/Lane.
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-30) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Western blot analysis of Phospho-ATF2 (T71) on NIH/3T3 cell lysates treated with or without untreated anisomycin(250ng/ml 30min) with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/1,000 dilution.
Lysates/proteins at 20 µg/Lane.
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-30) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of Hela cells treated with or without 250ng/mL anisomycin for 30 minutes labeling Phospho-ATF2 (T71) with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. -
ICC staining of Phospho-ATF2 (T71) in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-30, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
-
ICC staining of Phospho-ATF2 (T71) in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-30, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
-
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/800 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human prostate tissue using anti-Phospho-ATF2 (T71) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-Phospho-ATF2 (T71) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-Phospho-ATF2 (T71) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
-
Immunohistochemical analysis of paraffin-embedded human cervix tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human esophagus tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse large intestine tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/800 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Phospho-ATF2 (T71) was immunoprecipitated in 0.2mg HeLa cell treated anisomycin(25ug/ml 30min) lysate with (ET1610-30) at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using (ET1610-30) at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: (ET1610-30) IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of (ET1610-30) in HeLa cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 mintues
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
-
Jiawei Taohe Chengqi Decoction attenuates CCl4 induced hepatic fibrosis by inhibiting HSCs activation via TGF-β1/CUGBP1 and IFN-γ/Smad7 pathway
Journal: Phytomedicine
DOI: 10.1016/j.phymed.2024.155916
IF: 6.7
Application: WB
Reactivity: Mouse,Human
Publish date: 2024 Jul
Products with the same target and pathway
